Aquibacillus halophilus gen. nov., sp. nov., a moderately halophilic bacterium from a hypersaline lake, and reclassification of Virgibacillus koreensis as Aquibacillus koreensis comb. nov. and Virgibacillus albus as Aquibacillus albus comb. nov. A novel Gram-stain-positive, moderately halophilic bacterium, designated strain B6B T , was isolated from the water of an Iranian hypersaline lake, Aran-Bidgol, and characterized taxonomically using a polyphasic approach. Cells of strain B6B T were rod-shaped, motile and produced ellipsoidal endospores in terminal positions in non-swollen sporangia. Strain B6B T was a strictly aerobic bacterium and catalase-and oxidase-positive. The strain was able to grow at NaCl concentrations of 0.5-20.0 % (w/v), with optimum growth occurring at 10.0 % (w/v) NaCl. The optimum temperature and pH for growth were 35 6C and pH 7.0. On the basis of 16S rRNA gene sequence analysis, strain B6B T was shown to belong to the phylum Firmicutes and its closest phylogenetic similarities were with the species Virgibacillus koreensis BH30097 T (97.5 %), Virgibacillus albus YIM 93624 T (97.4 %), Sediminibacillus halophilus EN8d T (96.8 %), Sediminibacillus albus NHBX5 T (96.6 %), Virgibacillus carmonensis LMG 20964 T (96.3 %) and Paraliobacillus quinghaiensis YIM-C158 T (96.0 %), respectively. Phylogenetic analysis revealed that strain B6B T , along with V. koreensis BH30097 T and V. albus YIM 93624 T , clustered in a separate clade in the family Bacillaceae. The DNA G+C content of the novel isolate was 35.8 mol%. DNA-DNA hybridization experiments revealed low levels of relatedness between strain B6B T and V. koreensis BH30097 T (13 %) and V. albus YIM 93624 T (33 %). The major cellular fatty acid of strain B6B T was anteiso-C 15 : 0 (75.1 %) and its polar lipid pattern consisted of phosphatidylglycerol, diphosphatidylglycerol, an unknown phospholipid and an unknown glycolipid. The isoprenoid quinones were MK-7 (90 %) and MK-6 (3 %). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. All of these features support the placement of isolate B6B T within the phylum Firmicutes. It is closely related to V. koreensis and V. albus, but with features that clearly distinguish it from species of the genus Virgibacillus or of other related genera. On the basis of the polyphasic evidence derived in this study, we propose that strain B6B T be placed within a new genus, as Aquibacillus halophilus gen. nov., sp. nov., with B6B T as the type strain (5IBRC-M 10775 T 5KCTC 13828 T ). We also propose that V. koreensisOne supplementary table and three supplementary figures are available with the online version of this paper.The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain B6B T is HQ433456.

A novel Gram-stain-positive, moderately halophilic bacterium, designated strain E33T , was isolated from water of the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells of strain E33 T were motile rods and produced ellipsoidal endospores at a central or subterminal position in swollen sporangia. Strain E33 T was a strictly aerobic bacterium, catalase-and oxidase-positive. The strain was able to grow at NaCl concentrations of 0.5-25 % (w/v), with optimum growth occurring at 5-15 % (w/v) NaCl. The optimum temperature and pH for growth were 40 6C and pH 7.5-8.0, respectively. On the basis of 16S rRNA gene sequence analysis, strain E33 T was shown to belong to the genus Bacillus within the phylum Firmicutes and showed the closest phylogenetic similarity with the species Bacillus niabensis 4T19 T (99.2 %), Bacillus herbersteinensis D-1-5a T (97.3 %) and Bacillus litoralis SW-211 T (97.2 %). The DNA G+C content of the type strain of the novel species was 42.6 mol%. The major cellular fatty acids of strain E33 T were anteiso-C 15 : 0 and iso-C 15 : 0 , and the polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids, an unknown lipid and an unknown phospholipid. The isoprenoid quinones were MK-7 (97 %), MK-6 (2 %) and MK-8 (0.5 %). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. All these features confirm the placement of isolate E33 T within the

A Gram-positive, moderately halophilic rod, designated X5B T , was isolated from saline mud of the hypersaline lake Aran-Bidgol in Iran. Strain X5B T was a strictly aerobic, motile bacterium that produced ellipsoidal endospores at a central-subterminal position in non-swollen sporangia. The isolate grew at pH 7.0-10.0 (optimum pH 7.5), at 25-45 6C (optimum 35 6C) and with 2.5-15 % (w/v) NaCl (optimum 5-7.5 %). On the basis of 16S rRNA gene sequences, strain X5B T belonged to the genus Bacillus and showed highest similarity with Bacillus persepolensis HS136 T (95.6 % 16S rRNA gene sequence similarity) and Bacillus salarius BH169 T (95.5 %). The DNA G+C content was 42.4 mol%. The major cellular fatty acids were anteiso-C 15 : 0 and iso-C 15 : 0 and the polar lipid profile consisted of phosphatidylglycerol, diphosphatidylglycerol, three phospholipids and two glycolipids. The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid and the isoprenoid quinones were MK-7 (92 %), MK-6 (6 %) and MK-5 (2 %). On the basis of phylogenetic, chemotaxonomic and phenotypic data, a novel species of the genus Bacillus is proposed, with the name Bacillus iranensis sp. nov. The type strain is X5B T (5IBRC 10446 T 5DSM 23995 T ).Moderately halophilic bacteria are represented by physiologically and taxonomically heterogeneous groups that include both Gram-negative and Gram-positive microorganisms (Ventosa et al., 1998;Ventosa, 2006). Among the Gram-positive organisms, several aerobic, endosporeforming, halophilic genera and species have been reported (Arahal & Ventosa, 2002;Ventosa, 2006;de la Haba et al., 2011). These bacteria are frequently isolated from saline and hypersaline environments such as soils and aquatic habitats (Arahal & Ventosa, 2002;Ventosa, 2006; Márquez et al., 2011), are characterized by optimal growth in media containing 3-15 % (w/v) NaCl (Ventosa et al., 1998) and belong to species of the genus Bacillus or other Bacillusrelated genera (de la Haba et al., 2011). Studies of such bacteria are of great importance as they may produce compounds of industrial interest, such as extracellular hydrolytic enzymes that have diverse potential uses in biotechnological processes (Margesin & Schinner, 2001; Sánchez-Porro et al., 2003;Amoozegar et al., 2008;Kiran & Chandra, 2008;Rohban et al., 2009;Karbalaei-Heidari et al., 2009;Shafiei et al., 2010).During the study of the microbial population in hypersaline lakes in Iran, an aerobic, Gram-positive, moderately halophilic bacterium, designated strain X5B T , was isolated from mud of the hypersaline lake Aran-Bidgol, the largest playa in the centre of Iran, 1000 km distant from the coast and at an altitude of 800 m (sampling site: 34.64378 u N 51.83838 u E). This playa was formed by deposition of halite sediments in different geological periods. Most salts in this lake are NaCl, Na 2 SO 4 , MgCl 2 and MgSO 4 and there are traces of carbonate ions. The pH of the water of the lake is neutral (about pH 7.0-7.5) and its salinity reaches saturation. , 1982) ...

A novel Gram-positive, slightly halophilic bacterium, designated strain B48 T , was isolated from soil around the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells of strain B48T were non-motile rods and produced ellipsoidal endospores at a central or subterminal position in swollen sporangia. Strain B48 T was a strictly aerobic bacterium, catalase-and oxidase-positive. The strain was able to grow at NaCl concentrations of 0.5-10.0 % (w/v), with optimum growth occurring at 2.5 % (w/v) NaCl. The optimum temperature and pH for growth were 35 6C and pH 7.5-8.0, respectively. On the basis of 16S rRNA gene sequence analysis, strain B48 T was shown to belong to the genus Bacillus within the phylum Firmicutes and showed the closest phylogenetic similarity to the species Bacillus foraminis CV53 T (97.4 %) and Bacillus purgationiresistens DS22 T (96.9 %). The DNA G+C content of this new isolate was 40.1 mol%. The major cellular fatty acids of strain B48 T were iso-C 15 : 0 and anteiso-C 15 : 0 , and its polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an aminophospholipid and two unknown phospholipids. The only quinone present was menaquinone 7 (MK-7). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid.

A Gram-negative, non-endospore-forming, rod shaped, strictly aerobic, moderately halophilic bacterium, designated strain M9B(T), was isolated from the hypersaline lake Aran-Bidgol in Iran. Cells of strain M9B(T) were found to be motile and produce colonies with an orange-yellow pigment. Growth was determined to occur between 5 and 20 % (w/v) NaCl and the isolate grew optimally at 7.5-10 % (v/w) NaCl. The optimum pH and temperature for growth of the strain were determined to be pH 7.0 and 35 °C, respectively, while it was able to grow over pH and temperature ranges of 6-8 and 25-45 °C, respectively. Phylogenetic analysis based on the comparison of 16S rRNA gene sequences revealed that strain M9B(T) is a member of the genus Marinobacter. The closest relative to this strain was found to be Marinobacter hydrocarbonoclasticus MBIC 1303(T) with a similarity level of 97.7 %. DNA-DNA hybridization between the novel isolate and this phylogenetically related species was 13 ± 2 %. The major cellular fatty acids of the isolate were identified as C16:0, C19:1 ω6c, C18:1 ω9c and C16:1 ω9c. The polar lipid pattern of strain M9B(T) was determined to consist of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine and three phospholipids. Ubiquinone 9 (Q-9) was the only lipoquinone detected. The G+C content of the genomic DNA of this strain was determined to be 58.6 mol%. Phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data suggest that this strain represents a novel species of the genus Marinobacter, for which the name Marinobacter persicus sp. nov. is proposed. The type strain of Marinobacter persicus is strain M9B(T) (=IBRC-M 10445(T) = CCM 7970(T) = CECT 7991(T) = KCTC 23561(T)).